UPLC-Q-TOF-MS^E结合外源性代谢组学方法分析鉴定肝豆汤在大鼠尿液中的代谢产物  

Identification of major bioactive components and metabolites of Gandou decoction in rat urine by an integrative approach based on UPLC-Q-TOF-MS^E coupled with xenometabolomics analytical platform

作　　者：张雪燕 刘艺 吴欢 曹仕健[2] 吴鹏 周安 ZHANG Xue-yan;LIU Yi;WU Huan;CAO Shi-jian;WU Peng;ZHOU An(The Experimental Research Center,Anhui Province Key Laboratory of Chinese Medicinal Formula,Anhui University of Chinese Medicine,Hefei 230038,China;The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China;Waters Corporation(China),Shanghai 201206,China)

机构地区：[1]安徽中医药大学科研实验中心,中药复方安徽省重点实验室,安徽合肥230038 [2]安徽中医药大学第一附属医院,安徽合肥230031 [3]沃特世科技(上海)有限公司,上海201206

出　　处：《药学学报》2020年第5期971-978,共8页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金项目(81872976);安徽省重点研究与开发计划(201904a07020051);安徽省自然科学基金项目(1908085MH266);安徽省高校优秀青年骨干人才国外访学研修项目(gxgwfx2018042).

摘　　要：采用UPLC-Q-TOF-MS^E技术结合外源性代谢组学方法,对肝豆汤给药后正常和铜负荷模型大鼠尿液中的原形成分及代谢产物进行定性分析,在正、负离子扫描模式下获取大鼠尿液样品的质谱信息,包括保留时间、精确分子质量及离子强度。应用主成分分析(PCA)、正交偏最小二乘法-判别分析(OPLS-DA)外源性代谢组学方法筛选鉴定肝豆汤原形成分和相关代谢产物,最终从大鼠尿液中共检测到77个外源性成分,包括45个原形成分和32个代谢产物, 10个原形成分经对照品确证;肝豆汤在大鼠尿液中原形成分主要有蒽醌、生物碱、原萜烷型萜和黄酮;这些成分在大鼠尿液中代谢途径有羟基化、去甲基化、硫酸化、葡萄糖醛酸化等。结果表明UPLC-Q-TOF-MS^E结合外源性代谢组学方法准确便捷,可作为筛选鉴定体内微量代谢成分的有效工具。动物实验经安徽中医药大学实验动物伦理委员会批准(批准号:2019025)。To identify major bioactive components and metabolites of Gandou decoction(GDD) in urine of normal and copper-laden rats, an integrative approach that ultra-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS^E) coupled with xenometabolomics analytical platform was established. Mass spectral data information about retention time, accurate m/z and ionic strength of rat urine samples was performed under positive and negative ion modes. Unsupervised principal components analysis(PCA) and supervised orthogonal partial least-squared discriminant analysis(OPLS-DA) were used to reveal the differential ions. As a result, a total of 77 compounds including 45 prototypes and 32 metabolites in urine were detected. Results indicated that anthraquinones, alkaloids and tetracyclic triterpenoids and flavonoids were the main chemical components of GDD in rat urine;the main metabolic pathways of these compounds in rat urine mainly include hydroxyl, methylation, sulfating, glucuronidation, and so on. UPLC-QTOF-MS^E coupled with xenometabolomics analytical platform is fast and efficient so that facilitates authentication of the material basis of Chinese herb compound in vivo, can also be used as an effective tool for ascertaining trace bioactive components in vivo.The animal experiments were approved by the Experimental Animal Ethics Committee of Anhui University of Chinese Medicine(No. 2019025).

关 键 词：肝豆汤 外源性代谢组学 UPLC-Q-TOF-MS^E 威尔逊病 代谢产物 

分 类 号：R917[医药卫生—药物分析学]